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. 2013 Sep 25;8(9):e75472. doi: 10.1371/journal.pone.0075472

Figure 1. Strain 7254 is defined by a K915X mutation and generates a non-functional truncated Jak2 protein.

Figure 1

(A) The Jak2 domain structure is indicated. The DNA sequence from 7254 splenocytes and corresponding protein sequence are also shown. (B) Splenocytes were harvested from phenylhydrazine primed Jak2 K915X and Jak2 +/- mice and their wild type littermates. HA-tagged Jak2 and Jak2K915X were also expressed in 293T cells. A Western blot was performed with a peptide-specific JAK2 antibody. (C) 293T cells were transfected with cDNAs encoding HA-Jak2 or HA-Jak2 K915X. Western blotting was performed with phosphorylation-specific antibodies that recognize pSer-523, pTyr-570 and pTyr-1007/1008 in Jak2. The membranes were stripped and reprobed with an anti-HA antibody. (D) HA-tagged versions of Jak2 and Jak2K915X with or without the JAK2 3’ UTR were expressed in 293T cells. Western blots were performed with 4G10 anti-phosphotyrosine and HA antibodies. Immunoblotting with anti β-tubulin was performed to demonstrate equal loading.